Insulin like growth factor 1

Manufactured by Lonza

Sourced in United States, Switzerland

Insulin-like growth factor-1 (IGF-1) is a laboratory protein that plays a key role in cell growth and development. It is a critical component in various cell-based assays and research applications.

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Lab products found in correlation

Heparin, by Lonza (3 mentions) Hydrocortisone, by Lonza (3 mentions) Penicillin streptomycin solution, by Thermo Fisher Scientific (2 mentions) Recombinant human fb growth factor, by Lonza (2 mentions) Gentamicin amphotericin, by Lonza (2 mentions) Basic fibroblast growth factor (bfgf), by Lonza (2 mentions) Ascorbic acid, by Lonza (2 mentions) Epidermal growth factor (egf), by Lonza (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Lonza (2 mentions) Pcs 100 010, by American Type Culture Collection (1 mentions) Crl 1634, by American Type Culture Collection (1 mentions) Dmem high glucose, by Welgene (1 mentions) Hacat, by Addexbio (1 mentions) Vascular endothelial growth factor (vegf), by Lonza (1 mentions) Human epithelial growth factor, by Lonza (1 mentions) Pooled huvec, by Lonza (1 mentions) Egm 2 bulletkit, by Lonza (1 mentions) Gentamycin, by Lonza (1 mentions) Amphotericin b, by Lonza (1 mentions)

6 protocols using insulin like growth factor 1

Cell Culture Protocols for Skin Tissue Research

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We used HaCaT cells, HS27 fibroblasts (FBS), and HUVECs as substitutes for the epithelial cells, FBS and blood vessel endothelial cells present in skin tissue54 (link). The human epidermal keratinocytes (HaCaT, ATCC^®, USA) and human skin Fb HS27 cells (CRL-1634, ATCC^®, USA) were cultured in high-glucose DMEM (WELGENE, South Korea) supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY) and 1% penicillin/streptomycin solution (Gibco, Grand Island, NY). The HUVECs (PCS-100-010^™, ATCC^®, USA) were cultured in EGM-2 supplemented with ascorbic acid, vascular endothelial growth factor, 20% FBS, recombinant human Fb growth factor, hydrocortisone, insulin-like growth factor-1, a recombinant analog of human insulin-like growth factor-1, gentamicin–amphotericin, and heparin (Lonza, Walkersville, MD, USA). All experiments used cells from passages 3–10.

Wufuer M., Lee G., Hur W., Jeon B., Kim B.J., Choi T.H, & Lee S. (2016). Skin-on-a-chip model simulating inflammation, edema and drug-based treatment. Scientific Reports, 6, 37471.

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Culturing Skin Cell Lines for Research

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Human epidermal keratinocytes HaCaT (Addexbio, San Diego, CA, USA), a spontaneously transformed immortal human keratinocyte cell line, and human skin fibroblasts Hs27 (CRL1735, ATCC^®, Manassas, VA, USA) were cultured in high-glucose DMEM (WELGENE, Daegu, Korea) supplemented with 10% fetal bovine serum (Gibco, Grand Island, NY, USA) and 1% penicillin/streptomycin solution (Gibco). The HUVECs (PCS-100-01, ATCC^®) were cultured in EGM-2 supplemented with ascorbic acid, vascular endothelial growth factor, 20% FBS, recombinant human Fb growth factor, hydrocortisone, insulin-like growth factor-1, a recombinant analog of human insulin-like growth factor-1, gentamicin–amphotericin, and heparin (Lonza, Basel, Switzerland). HaCaT, Hs27, and HUVECs were used as substitutes for epithelial cells, fibroblasts, and blood vessel endothelial cells present in the skin tissue. All experiments used cells from passages 3–10. All cells were incubated in 5% CO₂ at 37 °C.

Choi D.H., Jeon B., Lim M.H., Lee D.H., Ye S.K., Jeong S.Y, & Kim S. (2021). 3D cell culture using a clinostat reproduces microgravity-induced skin changes. NPJ Microgravity, 7, 20.

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Endothelial Cell Culture and Characterization

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Pooled HUVEC (from 3 to 6 individual donors, Lonza, Cologne, Germany) and HUAEC (from 250 individual donors, PeloBiotech GmbH, Planegg, Germany) were cultivated in EGM-2 Bullet Kit containing endothelial cell culture medium supplemented with 2% (v/v) fetal bovine serum, vascular endothelial growth factor, basic fibroblast growth factor, human epithelial growth factor, insulin-like growth factor-1, hydrocortisone, heparin, ascorbic acid, gentamycin, and amphotericin B (Lonza) in a standard humidified incubator at 37 °C with 5% (v/v) CO₂. Cells of passage 5 were seeded in 4-well PCA chamber slides (Sarstedt, Nürnbrecht, Germany) at a cell density of 15,000 cells/well with 1 mL medium per well, and cultivated for up to 7 days. Medium was exchanged at day 2 and day 4 of cultivation.

Lau S., Gossen M., Lendlein A, & Jung F. (2021). Venous and Arterial Endothelial Cells from Human Umbilical Cords: Potential Cell Sources for Cardiovascular Research. International Journal of Molecular Sciences, 22(2), 978.

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Isolation and Characterization of Endothelial Outgrowth Cells

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Mouse EOCs were cultured as previously described [19 (link)]. Briefly, macrophage-depleted BMMNCs were seeded in fibronectin-coated plates and maintained in endothelial cell basal medium-2 supplemented with 5% FBS, VEGF-A, fibroblast growth factor, insulin-like growth factor-1, epidermal growth factor, ascorbic acid and antibiotics (Lonza, Walkersville, MD, USA). Non-adherent cells were removed after 4 days of culture and new medium was applied. EOCs, recognised as an attached cluster of spindle-shaped cells [20 (link)], were characterised on day 7 by immunostaining using primary antibodies against CD34, CD31, VEGFR2 and VE-cadherin (ESM Table 1). After incubation with Alexa Flour 488- or 594-conjugated secondary antibodies (Invitrogen, Carlsbad, CA, USA), the cells were visualised under an Olympus BX53 microscope (Olympus, Tokyo, Japan).

Bhatta M., Ma J.H., Wang J.J., Sakowski J, & Zhang S.X. (2015). Enhanced endoplasmic reticulum stress in bone marrow angiogenic progenitor cells in a mouse model of long-term experimental type 2 diabetes. Diabetologia, 58(9), 2181-2190.

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Aortic Ring Angiogenesis Assay

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The aortic ring assay was performed as previously described.³³ (link) In brief, mouse thoracic aortas were isolated and cut into 1-mm-wide rings and embedded in Matrigel (Corning, Corning, NY) and incubated in EBM-2 (Lonza Inc, Basel, Switzerland) supplemented with VEGFA alone or full EC growth medium-2 (EGM-2) containing VEGFA, fibroblast growth factor, epidermal cell growth factor, and insulin-like growth factor-1 (Lonza Inc). The media were replaced every 2 days. For each sample, the length of eight sprouts at days 6 and 8, originating from the aorta, were quantified using the Fiji software (available at: http://fiji.sc/Fiji) from brightfield images taken at 2× magnification.

Kiesworo K., MacArthur M.R., Kip P., Agius T., Macabrey D., Lambelet M., Hamard L., Ozaki C.K., Mitchell J.R., Déglise S., Mitchell S.J., Allagnat F, & Longchamp A. (2023). Cystathionine-γ-lyase overexpression modulates oxidized nicotinamide adenine dinucleotide biosynthesis and enhances neovascularization. JVS-Vascular Science, 4, 100095.

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Endothelial Cell Differentiation Induction

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HRMEC were purchased from Cell Systems (Kirkland, WA) and cultured as described previously (Nawaz et al., 2013) in endothelial basal medium-2 (EBM-2) enriched with endothelial growth medium-2 MV Bulletkit, containing, besides fetal calf serum [FCS; final concentration 5% (v/v)], the growth factors epidermal growth factor, vascular endothelial growth factor (VEGF), basic fibroblast growth factor and insulin-like growth factor 1 (Lonza, Verviers, Belgium). Three days after seeding in 6-well plates, differentiation of HRMEC was induced by addition of human recombinant IL-1, human recombinant TNF-, human recombinant TGF-1 or human recombinant connective tissue growth factor (CTGF) (all from PeproTech, Rocky Hill, NJ) or combinations thereof and replacement of the growth-factor enriched endothelial cell growth medium by EBM-2 supplemented with 3% FCS. As a control to preserve the endothelial cell type, cultures in one well of each plate were contained in the endothelial cell growth medium supplemented with 10 ng/ml of VEGF. The differentiation medium was replaced every two days. After 4 days of stimulation, cell morphology and gene expression were investigated. Changes in cell morphology were documented using an Axiovert 200M inverted microscope, equipped with an EC Plan-Neofluar 10× phase contrast objective (Carl Zeiss, Jena, Germany).

Abu El-Asrar A.M., De Hertogh G., van den Eynde K., Alam K., Van Raemdonck K., Opdenakker G., Van Damme J., Geboes K, & Struyf S. (2015). Myofibroblasts in proliferative diabetic retinopathy can originate from infiltrating fibrocytes and through endothelial-to-mesenchymal transition (EndoMT). Experimental eye research, 132.

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